Abstract: Traditional Au spheres (AuSPs) based test strips cannot meet the requirements of high-sensitive detection of food safety due to their low detection sensitivity. Therefore, it is particularly important to establish highly sensitive and rapid analysis methods. With aflatoxin B₁ (aflatoxin B₁, AFB₁) as the detection object, Au nanorods (AuNRs) as the signal tag, conjugated anti-AFB₁ monoclonal antibody as the gold labeled probe, immunochromatographic test strip were prepared based on competition method for sensitive detection of AFB₁ in maize. The experimental results were quantified by a portable test strip quantitative analyzer. The standard curve was prepared under the conditions of 0.6 μg monoclonal antibody addition, 0.1 mg/mL T-line antigen concentration and 2.5 μL probe usage volume, the detection limit of the strip based on AuNRs was 0.124 ng/mL, IC₅₀ was 1.593 ng/mL. In addition, the recovery rate of AFB₁ in the actual maize samples was 89.5% to 111.70%. The maize quality control samples were tested, and the results were consistent with the established method. The method can be applied to the rapid detection of AFB₁ in maize, and the sample detection results meet the national standard and have a good application prospect.